Journal: Nature Communications

Article Title: Nanoparticles targeting mutant p53 overcome chemoresistance and tumor recurrence in non-small cell lung cancer

doi: 10.1038/s41467-024-47080-3

Figure Lengend Snippet: a Kaplan-Meier plot of the correlation between the mutation of p53 and the survival of patients with NSCLC (Log-rank Mantel–Cox test). Cisplatin (Cis), fluvastatin (Flu). b Outline of the assays of cisplatin treatment (low dose, 10 μM; moderate (mod) dose, 25 μM; high dose, 50 μM). Cholesterol (Cho). Heatmap of the number of mutations and RNA-seq analysis ( c ), ROS levels ( d ), Cho levels ( e ), and DNA damage ( f ) for wtp53-expressing cells treated with cisplatin for 30 days. g High-throughput sequencing before and after fluvastatin sodium (4 μM) treatment in A549 cells. h Genome-wide analysis. The volcano plot depicts the significance and magnitude of difference (Fold Change). The dashed line indicates the threshold of the Fold Change > 2 and adjusted p < 0.05. Some of the cancer related genes are labeled by dark colors. i GO enrichment analysis of differentially expressed genes (DEGs). The advanced bubble chart shows GO enrichment of DEGs in signaling pathways. The x-axis label represents the gene ratio, and the y-axis label represents GO terms. j GSEA analysis. The normalized enrichment scores (NES) and p values are indicated in each plot. k Heatmap analysis of mevalonate pathway genes from RNA-seq data. The color scale indicates the fold change in genes expression. l Schematic illustration of treatment with cisplatin. b , g created with BioRender.com. Data are shown as the mean ± SD; n.s. = no significance. Source data are provided as a file.

Article Snippet: The p53 mutant human NSCLC cell lines NCI-H1975 (American Type Culture Collection (ATCC), #CRL-5908), NCI-H2087 (ATCC, #CRL-5922), NCI-H2342 (ATCC, #CRL-5941), NCI-H1793 (ATCC, #CRL-5896), NCI-H2196 (ATCC, #CRL-5932), NCI-H748 (ATCC, #CRL-5841), NCI-H1770 (ATCC, #CRL-5893), NCI-H1591 (ATCC, #CRL-5878), and NCI-H520 (ATCC, #HTB-182) were purchased from ATCC.

Techniques: Mutagenesis, RNA Sequencing, Expressing, Next-Generation Sequencing, Genome Wide, Labeling, Protein-Protein interactions

Journal: Nature Communications

Article Title: Nanoparticles targeting mutant p53 overcome chemoresistance and tumor recurrence in non-small cell lung cancer

doi: 10.1038/s41467-024-47080-3

Figure Lengend Snippet: a Schematic summary of the p53-independent antitumor mechanism of FP NPs. b Confocal images of Dil and ER-Tracker in H1975 cells treated with 2 μM Dil@FP NPs. Their colocalization determined by Pearson’s correlation coefficient was quantified ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Scale bars, 10 μm. c Confocal images of Dil and Fluo-4 in H1975 cells treated with 2 μM Dil@FP NPs. Their fluorescence intensity was quantified ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Scale bars, 10 μm. d Confocal images of Dil and MitoTracker in H1975 cells treated with 2 μM Dil@FP NPs, and their colocalization determined by Pearson’s correlation coefficient was quantified ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Scale bars, 10 μm. e Confocal images of JC-1 in H1975 cells treated with 4 μM different formulations (dosage based on Fluplatin) for 6 h, and their fluorescence intensity was quantified ( j ), ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Scale bars, 10 μm. Western blotting analysis of p-elF2α, elF2α, CHOP, and ATF4 in H1975 cells ( f ) and A549 cells ( g ) after treatment with 4 μM of different formulations (dosage based on Fluplatin; n = 3 independent samples). h Western blotting analysis of p-elF2α, elF2α, CHOP, and ATF4 in H1975 cells after treatment with different concentration of FP NPs for 12 h ( n = 3 independent samples). i Confocal images of IF staining against γ-H2AX in H1975 cells treated with 4 μM FP NPs for 6 h, and their fluorescence intensity was quantified ( k ), ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Scale bars, 10 μm. a Created with BioRender.com. Data are shown as the mean ± SD; n.s. no significance. Source data are provided as a file.

Article Snippet: The p53 mutant human NSCLC cell lines NCI-H1975 (American Type Culture Collection (ATCC), #CRL-5908), NCI-H2087 (ATCC, #CRL-5922), NCI-H2342 (ATCC, #CRL-5941), NCI-H1793 (ATCC, #CRL-5896), NCI-H2196 (ATCC, #CRL-5932), NCI-H748 (ATCC, #CRL-5841), NCI-H1770 (ATCC, #CRL-5893), NCI-H1591 (ATCC, #CRL-5878), and NCI-H520 (ATCC, #HTB-182) were purchased from ATCC.

Techniques: Fluorescence, Western Blot, Concentration Assay, Staining

Journal: Nature Communications

Article Title: Nanoparticles targeting mutant p53 overcome chemoresistance and tumor recurrence in non-small cell lung cancer

doi: 10.1038/s41467-024-47080-3

Figure Lengend Snippet: a Western blotting analysis of p53 in cells after treatment with 4 μM of different formulations (dosage based on Fluplatin) for 12 h. Their grayscale values were quantified ( b – e ) ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). f Western blotting analysis of p53 in cells after treatment with different concentrations of FP NPs for 12 h. Their grayscale values were quantified ( g , h ) ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). i Western blotting analysis of p53 in cells after treatment with 4 μM of FP NPs for 12 h. And their grayscale values were quantified ( j ) ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). k Western blot of cells treated with 4 μM FP NPs or not for 12 h and cultured in medium containing 10 μM MG132 or 10 mM 3-MA. Their grayscale values were quantified ( l ) ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). Western blot of H1975 ( m ), H2087 ( n ), H2342 ( o ), and A549 ( p ) cells treated with 2 μM FP NPs or not for 12 h and then treated with 100 μg/mL CHX at the indicated time points. n = 3 biologically independent samples, relative p53/actin ratios are shown. q Western blotting analysis of total ubiquitination (Ub) of immunoprecipitated (IP) p53 in cells after treatment with 2 μM FP NPs with or without MG132 ( n = 3 independent samples). r HMGR inhibition assay under different formulations ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). s The volcano plot shows differential expression of FP NPs treated and untreated H1975 cells. t Schematic summary of the p53-related antitumor mechanism of FP NPs. Data are shown as the mean ± SD; n.s. no significance. Source data are provided as a file.

Article Snippet: The p53 mutant human NSCLC cell lines NCI-H1975 (American Type Culture Collection (ATCC), #CRL-5908), NCI-H2087 (ATCC, #CRL-5922), NCI-H2342 (ATCC, #CRL-5941), NCI-H1793 (ATCC, #CRL-5896), NCI-H2196 (ATCC, #CRL-5932), NCI-H748 (ATCC, #CRL-5841), NCI-H1770 (ATCC, #CRL-5893), NCI-H1591 (ATCC, #CRL-5878), and NCI-H520 (ATCC, #HTB-182) were purchased from ATCC.

Techniques: Western Blot, Cell Culture, Ubiquitin Proteomics, Immunoprecipitation, Inhibition, Quantitative Proteomics

Journal: Nature Communications

Article Title: Nanoparticles targeting mutant p53 overcome chemoresistance and tumor recurrence in non-small cell lung cancer

doi: 10.1038/s41467-024-47080-3

Figure Lengend Snippet: a Ex vivo biodistribution imaging of the main organs in mice bearing H1975 xenografts of the DiR@FP NPs at various time points post injection. Scale bars, 5 mm. b ICP‒MS measurement of Pt accumulation in individual organs at 1, 6, 12, and 24 h after treatment with cisplatin (i) or FP NPs (ii) ( n = 3 mice per group). c ICP‒MS measurement of Pt accumulation in the tumor tissues at 1, 6, 12, and 24 h after treatment with cisplatin or FP NPs ( n = 3 mice per group; one-way ANOVA followed by Tukey’s HSD post hoc test). d TC levels in tumor tissues and TC, TG, HDL, LDL levels in serum after treatment with each group ( n = 3 mice per group; one-way ANOVA followed by Tukey’s HSD post hoc test). Confocal images of ATF4 ( e ) and γ-H2AX ( f ) in the tumor tissues of mice after treatment with each group. Scale bars, 100 μm. g Western blotting analysis of p53 R273H in tumor tissues after treatment with each group. Their grayscale values were quantified ( h ) ( n = 3 independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). i IHC of p53 in the tumor tissues after treatment with each group. Scale bars, 20 μm. j Kaplan-Meier survival curve of mice treated with each group over 70 days ( n = 8 mice per group; Log-rank Mantel–Cox test). k The body weight of mice during treatments with each group over 70 days ( n = 8 mice per group;two-way ANOVA followed by Tukey’s multiple comparisons post test). l IHC of Ki67 in the tumor tissues after treatment with each group. Scale bars, 20 μm. Data are shown as the mean ± SD; n.s. no significance. Source data are provided as a file.

Article Snippet: The p53 mutant human NSCLC cell lines NCI-H1975 (American Type Culture Collection (ATCC), #CRL-5908), NCI-H2087 (ATCC, #CRL-5922), NCI-H2342 (ATCC, #CRL-5941), NCI-H1793 (ATCC, #CRL-5896), NCI-H2196 (ATCC, #CRL-5932), NCI-H748 (ATCC, #CRL-5841), NCI-H1770 (ATCC, #CRL-5893), NCI-H1591 (ATCC, #CRL-5878), and NCI-H520 (ATCC, #HTB-182) were purchased from ATCC.

Techniques: Ex Vivo, Imaging, Injection, Western Blot

Journal: Nature Communications

Article Title: Nanoparticles targeting mutant p53 overcome chemoresistance and tumor recurrence in non-small cell lung cancer

doi: 10.1038/s41467-024-47080-3

Figure Lengend Snippet: a Schematic illustration of the experimental design. b The tumor volumes of mice during treatments with each group ( n = 7 mice per group; two-way ANOVA followed by Tukey’s multiple comparisons post test). c The body weight of mice during treatments with each group ( n = 7 mice per group; two-way ANOVA followed by Tukey’s multiple comparisons post test). d Tumor volume changes for each mouse in each group ( n = 7 mice per group). e In vivo bioluminescence imaging of tumor-bearing mice receiving various treatments after surgery. Three representative mice in each treatment group are shown. Images of day 0 were taken on the day of surgery. f The final anatomical picture and H&E staining of the lungs ( n = 3 independent samples; Scale bars, 5 mm). g IHC of p53, E-cadherin, MMP-2, MMP-9, Vimentin in the primary tumor tissues of cisplatin or FP NPs treatment, and recurrent and metastatic tumor tissues of cisplatin treatment ( n = 3 independent samples). Scale bars, 20 μm. h Recurrence tumor weights of different groups on day 42 after surgery ( n = 7 mice per group; one-way ANOVA followed by Tukey’s HSD post hoc test). i Kaplan-Meier survival curve of mice treated with each group over 65 days ( n = 7 mice per group; Log-rank Mantel–Cox test). j Final weights of the heart, liver, spleen, lungs, and kidneys ( n = 7 mice per group;two-tailed unpaired t test). k Heatmap of TNF-α, VEGF, IL-10 and IL-6 expression profiles in serum ( n = 3 mice per group). l Indicators of routine blood examination ( n = 3 mice per group; two-tailed unpaired t test) of mice. Data are shown as the mean ± SD; n.s. no significance. Source data are provided as a file.

Article Snippet: The p53 mutant human NSCLC cell lines NCI-H1975 (American Type Culture Collection (ATCC), #CRL-5908), NCI-H2087 (ATCC, #CRL-5922), NCI-H2342 (ATCC, #CRL-5941), NCI-H1793 (ATCC, #CRL-5896), NCI-H2196 (ATCC, #CRL-5932), NCI-H748 (ATCC, #CRL-5841), NCI-H1770 (ATCC, #CRL-5893), NCI-H1591 (ATCC, #CRL-5878), and NCI-H520 (ATCC, #HTB-182) were purchased from ATCC.

Techniques: In Vivo, Imaging, Staining, Two Tailed Test, Expressing

Journal: Plant Biotechnology Journal

Article Title: Affordable oral health care: dental biofilm disruption using chloroplast made enzymes with chewing gum delivery

doi: 10.1111/pbi.13643

Figure Lengend Snippet: Generation of Marker‐free (MF) lettuce plants expressing dextranase, mutanase and lipase and evaluation of transgene integration, marker removal and homoplasmy. Schematic representation of the integration of two expression cassettes (gene of interest—GOI and selectable marker) into lettuce chloroplast genome via homologous recombination of flanking sequences: 16S rRNA‐trnI and tranA‐23S rRNA and subsequent removal of the antibiotic resistance gene via homologous recombination between two identical atpB regions. GOI represents PG1‐Smdex or mut or lipY . Probe indicates the DNA fragment region which was digested by Bam HI and used to detect hybridizing fragments in Southern blots. (a) Southern blots confirm PG1‐Smdex gene integration, marker removal and homoplasmy in transplastomic plants with 10.5 kb with 2.2 kb fragments, while 12.5 kb with 10.5 kb and 2.2 kb demonstrated heteroplasmy (with or without the aadA gene) after gDNA was digested with Hind III. Untransformed plant (WT) and undigested DNA (UD) were used as controls (b). In MF mutanase T0 generation, expected bands of 14.1 or 16.1 kb as a result of Hind III digested gDNA confirm mut gene integration in T0 generation plants, and the 14.1 kb band alone represents the homoplasmy (c). Expected band size of 5.6 kb obtained from Sma I digested gDNA confirms lipY gene integration, antibiotic marker gene removal and homoplasmy in lipase expressing T1 generation plants (d). Gene of interest band size is represented with arrowheads.

Article Snippet: The Smdex gene (2574 bp, gene designation from Kim et al ., ) encoding dextranase was isolated from S. mutans strain ATCC 25175 genomic DNA using PCR (Figure A,B) and fused to the PG1 (encoding antimicrobial peptide Protegrin‐1).

Techniques: Marker, Expressing, Homologous Recombination

Journal: Plant Biotechnology Journal

Article Title: Affordable oral health care: dental biofilm disruption using chloroplast made enzymes with chewing gum delivery

doi: 10.1111/pbi.13643

Figure Lengend Snippet: Antibiofilm effects of commercial and plant‐derived enzymes against bacterial‐fungal mixed biofilms. Commercial purified enzymes of the same activity unit as measured in the plant crude extracts (333.3 U/mL for lipase and 7.08/0.84 U/mL for dextranase/mutanase, respectively) were used as standards to evaluate the antibiofilm efficacy of the plant crude extracts. (a) Confocal images showing the antibiofilm efficacy of commercial and plant‐derived lipase. (b–e) Quantitative computational analysis of the confocal biofilm images treated with commercial and plant‐derived lipase. (f) Confocal images showing the antibiofilm efficacy of commercial and plant‐derived Dextranase/Mutanase combination. (g–j) Quantitative computational analysis of the confocal biofilm images treated with commercial and plant‐derived Dextranase/Mutanase combination. For multi‐channel confocal images, C. albicans cells (yeasts or hyphae) are depicted in cyan; S. mutans cells are depicted in green; The EPS glucan matrix is depicted in red. For the computational data, the title of each graph indicates the channel(s) used for individual analysis. *, P < 0.05; **, P < 0.01 (one‐way analysis of variance with Tukey’s multiple comparisons test). Enzyme units of lipase and dextranase/mutanase represent µmol of pNP and reducing sugar produced in 1 h, respectively.

Article Snippet: The Smdex gene (2574 bp, gene designation from Kim et al ., ) encoding dextranase was isolated from S. mutans strain ATCC 25175 genomic DNA using PCR (Figure A,B) and fused to the PG1 (encoding antimicrobial peptide Protegrin‐1).

Techniques: Derivative Assay, Purification, Activity Assay, Produced

Journal: Plant Biotechnology Journal

Article Title: Affordable oral health care: dental biofilm disruption using chloroplast made enzymes with chewing gum delivery

doi: 10.1111/pbi.13643

Figure Lengend Snippet: Prevention of fungal‐bacterial mixed biofilm by topical sequential treatment of commercial Lipase and Dextranse/Mutanse combination. Commercial enzymes of the optimum activity units for bioactivty (1000 U/mL for lipase and 525/105 U/mL for dextranase/mutanase, respectively) were used. (a) Three‐dimensional confocal images of the fungal‐bacterial mixed biofilm formed after the topical sequential treatments. C. albicans cells (yeasts or hyphae) are depicted in cyan; S. mutans cells are depicted in green; The EPS glucan matrix is depicted in red. Representative merged biofilm images are displayed in the middle panel, while a magnified (close‐up) view of each small box is positioned in the left panel. Lateral (side) views of each biofilm are displayed at the right panel (the merged image at the top and the EPS channel at the bottom). (b–e) Quantitative computational analysis of the confocal images. The title of each graph indicates the channel(s) used for individual analysis. * P < 0.05; ** P < 0.01 (one‐way analysis of variance with Tukey’s multiple comparisons test). Enzyme unit of lipase and dextranase/mutanase represent µmol of pNP and reducing sugar produced in 1 h, respectively.

Article Snippet: The Smdex gene (2574 bp, gene designation from Kim et al ., ) encoding dextranase was isolated from S. mutans strain ATCC 25175 genomic DNA using PCR (Figure A,B) and fused to the PG1 (encoding antimicrobial peptide Protegrin‐1).

Techniques: Activity Assay, Produced